The detection and analysis of rare cells is a pervasive challenge in cellular biology and clinical diagnostics. Fluxion Biosciences proposes the development of a high throughput instrument and microfluidic consumable to conduct rare cell isolations for research and clinical applications. Targeted cell isolations are important to numerous fields such as cancer research, stem cell biology, prenatal care and immunological disorders. Quantifying the number of circulating tumor cells in the bloodstream is one such application that can lead to early cancer detection and can help monitor disease progression during treatment. These cells occur as low as one in ten million cells. Other compelling applications requiring rare cell detection include screening fetal cells during pregnancy for genetic aberrations and performing CD4 counts for immunocompromised patients with HIV. Modern laboratory tools such as centrifuges, cell sorters and cytometers are poorly equipped to detect and analyze cells appearing in such low frequencies. Although many kits are commercially available for conducting routine cell separations, most exhibit very poor recovery and purity yields when it comes to low prevalence target cells. This proposal aims to develop a microfluidic chip for rare cell isolations using the Company's well-plate microfluidic technology. The proposed research entails use of laminar flow streams to create efficient and accurate cell separations with high recovery and purity of targeted cells. The resultant product will deliver an automated, high throughput platform for conducting rare cell isolations on the research and clinical settings. Fluxion's approach represents a significant breakthrough over existing technologies which are mainly confined to low throughput, low efficiency cell isolations performed in filtration columns or well plates. The proposed system also enables multiple downstream analysis options with minimal cell handling and trauma. Cells will be available directly in the microfluidic devices for microscopic observation, high content screening and automated detection using a fluorescence plate reader. Cells will be easily removable from the devices for further analysis such as RT-PCR. Clinicians and researchers will be able to leverage this versatility of analysis and high throughput, high efficiency cell sorting to greatly enhance their productivity in life saving discoveries, diagnoses and treatments. PUBLIC HEALTH RELEVANCE: An unfortunate aspect of cellular biology is that the cells which hold the most promise for clinical diagnosis, disease progression and biological understanding often occur in the lowest frequencies. Circulating tumor cells can aid in the early detection and treatment of cancer yet appear as only one in ten million cells. A similar low prevalence is true for fetal cells found in the maternal bloodstream which could offer noninvasive ways to screen for genetic disorders earlier in pregnancy. Modern laboratory tools such as centrifuges, cell sorters and cytometers are simply not equipped to detect and analyze cells appearing in such low frequencies. This proposal aims to develop a high throughput system to isolate rare cells from biological samples and provide a range of analysis options to accommodate the desired endpoints of both clinical and research applications.